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Identification of a Distinctive Mutation Spectrum Associated with High Levels of Transcription in Yeast

机译：与酵母中高水平转录相关的独特突变谱的鉴定

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High levels of transcription are associated with increased mutation rates in Saccharomyces cerevisiae, a phenomenon termed transcription-associated mutation (TAM). To obtain insight into the mechanism of TAM, we obtained LYS2 forward mutation spectra under low- versus high-transcription conditions in which LYS2 was expressed from either the low-level pLYS2 promoter or the strong pGAL1-10 promoter, respectively. Because of the large size of the LYS2 locus, forward mutations first were mapped to specific LYS2 subregions, and then those mutations that occurred within a defined 736-bp target region were sequenced. In the low-transcription strain base substitutions comprised the majority (64%) of mutations, whereas short insertion-deletion mutations predominated (56%) in the high-transcription strain. Most notably, deletions of 2 nucleotides (nt) comprised 21% of the mutations in the high-transcription strain, and these events occurred predominantly at 5′-(G/C)AAA-3′ sites. No ?2 events were present in the low-transcription spectrum, thus identifying 2-nt deletions as a unique mutational signature for TAM.
机译：高水平的转录与啤酒酵母的突变率增加有关，该现象被称为转录相关突变（TAM）。为了深入了解TAM的机制，我们获得了在低转录与高转录条件下 LYS2 从低水平表达的 LYS2 正向突变谱> pLYS2 启动子或强 pGAL1-10 启动子。由于 LYS2 基因座的大小较大，因此先将正向突变定位到特定的 LYS2 子区域，然后对在定义的736-bp目标区域内发生的那些突变进行测序。在低转录菌株中，碱基取代占突变的大部分（64％），而在高转录菌株中，短插入-缺失突变占主导（56％）。最值得注意的是，2个核苷酸的缺失（nt）构成了高转录菌株中21％的突变，这些事件主要发生在5'-（G / C）AAA-3'位点。在低转录谱中不存在β2事件，因此将2-nt缺失鉴定为TAM的独特突变特征。

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